Thrombin On The Influence Of Synaptic Plasticity After Cerebral Hemorrhage

Objective:Intracerebral hemorrhage(ICH)is a common neurological disease with high mortality and morbidity,which may largely impact the patients' quality of life.Synaptic plasticity is closely related to neural functional recovery and can be adjust after ICH for quite a long time,which can reduce morbidity and improve the patients' quality of life.Thrombin is one of the series of serine protein hydrolase and has been studied in various fields.in recent years the Scientists found that thrombin can regulate synaptic plasticity and then promote neural functional recovery under both physiological and pathological conditions.In this experiment by hirudin restraining the activity of thrombin and by SCH79797 restraining the activity of PAR1 after ICH.,evaluate each group nerve function score and observe synaptic protein such as SYP and PSD-95 protein changes in the situation around hematoma and analyze SYP,PSD-95 protein content to explore thrombin on the influence of synaptic plasticity and its possible mechanism to provide theoretical basis for clinical treatment after cerebral hemorrhage.Methods: 1.using stereotactic apparatus,Autologous blood was injected into the caudate nucleus(fontanelle before the 0.2 mm,near the center line on the right side 3.0 mm)of rat that produced ICH model.after two hours animals awake use Zea-longa score grading standard score to slect score between 1 to 3 points of SD rats as model rats.2.Grouping and processing: The100 animals randomly ware divided into to each group,,sham operation group(Sham group),intracerebral hemorrhage(ICH group),cerebral hemorrhage group+ small dose hirudin group(SH group),cerebral hemorrhage + large dose hirudin group(LH group),ICH+PAR-1 inhibitor group(PI group)then they were divided into 3d,7 d,14 d,and 21 d four subgroups after operation,each subgroup five rats.Experimental animals for each category in the following treatment(1)sham operation group injecte 50 ul +10ul saline to the right caudate nucleus and 2 ml saline to the intraperitoneal;(2)ICH group injecte 50 ul autologous blood + 10 ul saline into the right caudate nucleus + 2 ml saline into the intraperitoneal;(3)SH group injecte 50 ul autologous blood +Hirudin 3 u(soluble in 10 u l saline)into the right caudate nucleusand and 2 ml saline into the intraperitoneal;(4)LH group injecte 50 ul autologous blood + Hirudin 5u(soluble in 10 u l saline)into the right caudate nucleusand and 2 ml saline into the intraperitoneal;(5)PI group,injecte 50 ul autologous blood + 10 ul saline into the right caudate nucleusand and 2ml SCH79797(25ug/kg Dissolved in 2 ml saline)into the intraperitonea.3.Specimen extraction and detection,3 d,7 d,point of experimental animals directly executed,14 d,21 d point of experimental animals first nerve function score and then let them dead.At corresponding time points anesthesia in rats,and perfusion,fixed in the brain,brain tissue slices.With Garcia nerve function score method to evaluate nerve function defect the score,The higher scores show the lighter neural dysfunction,the lower the score,the heavier neural function damage then put to death;with HE staining observation of brain tissue around hematoma basic morphology and structure;using immunohistochemical method to detect the time SYP and PSD-95 protein content around the hematoma in the five groups at different times around hematoma changed by Immunohistochemical and analyzed its average integral optical density.Results : 1.Each groups HE staining around the hematoma showed:(1)the Sham group,without brain hematoma,brain tissue structure at each time point is complete;(2)ICH group 3 d brain tissue around hematoma is a large number of blood cells,brain structure disorder,a large number of inflammatory cell infiltration,edema,severe nerve cells degeneration necrosis,even when the 7 d,14 d visible blood cells around the hematoma was gradually reduced,compared with 3 d,the number of inflammatory cells gradually reduce,continuously reduce edema degree and the nerve cells,to 21 d around the hematoma blood cells less even disappear almost disappeared,inflammatory cells,a.large number of fibrous tissue and glial cell hyperplasia,organization structure restoration of damaged.Compared with ICH group,SH group,LH group,each point in time the degree of inflammatory cell infiltration,edema and nerve cell degeneration necrosis is mitigated,damaged tissue repair is better than ICH grounp.what's the more SH group is the least amount of inflammatory cells infiltration and the lightest cell edema degree and degree of damaged tissue repair is the best.with ICH group,PI group each point in time the degree of inflammatory cell infiltration,edema and nerve cell degeneration necrosis is more serious,damaged tissue repair is a bad group.2.14 d,21d the nerve dysfunction score :(1)the Sham group without neurologic deficits(P>0.05),nerve function score the highest,while ICH group,SH group,LH group,PI group has a different degree of neurologic deficits and lower than Sham group(P < 0.05).there is also 14 d of the score was lower than those of 21 d(P < 0.05);(2)each time point,the SH group,LH group is higher than ICH group and SH group is higher than LH group statistically difference(P < 0.05);PI nerve function score below ICH group(P< 0.05).;3.The brain tissue around hematoma SYP protein content:(1)Sham group,only a small amount of SYP protein expression were positive and each point in time no significant difference(P > 0.05).(2)3 d,ICH group,SH group,LH group,PI group,only a small amount of SYP protein expression were positive,compared with the Sham group no significant difference(P > 0.05);7 d,14 d and 21 d,ICH group,SH group,LH group,PI groups of SYP positive protein expression significantly more than the Sham group(P < 0.05).(3)7 d,14 d and 21 d point in time,SH,LH group obviously higher than ICH group positive protein expression,while SH group was obviously higher than that of LH group are statistically difference(P < 0.05);7 d,14 d and 21 d point in time,PI SYP positive protein expression below ICH group(P < 0.05);(4)ICH group,SH group,LH,PI group each group from 7 d SYP positive protein increased,14 d to spike(P < 0.05),and then express to drop,but still at 21 d have more expression;4.The immunohistochemical display PSD-95 protein around hematoma(1)Sham group,only a small amount of PSD-95 protein expression were positive and each point in time no significant difference(P > 0.05).(2)3 d,ICH group,SH group,LH group,PI group,only a small amount of PSD-95 protein expression were positive,compared with the Sham group no significant difference(P > 0.05);7 d,14 d and 21 d,ICH group,SH group,LH group,PI groups of PSD-95 positive protein expression significantly more than the Sham group(P < 0.05).(3)7 d,14 d and 21 d point in time,SH,LH group obviously higher than ICH group positive protein expression,while SH group was obviously higher than that of LH group are statistically difference(P < 0.05);7 d,14 d and 21 d point in time PI group PSD-95 positive protein expression below ICH group(P < 0.05);(4)ICH group,SH group,LH,PI group each group from 7 d PSD-95 positive protein increased,14 d to spike(P < 0.05),and then express to drop,but still at 21 d have more expression;5.Linear correlation about 14 d SYP and PSD-95 protein expression and nerve dysfunction analysis :(1)among ICH group,SH group,LH group,PI group,SYP protein expression is positively associated with nerve dysfunction(r =0.948,P<0.01);(2)among ICH group,SH group,LH group,PI group,PSD-95 protein expression is positively associated with nerve dysfunction(r=0.925,P<0.001);Conclusion:1.using no anticoagulant autologous blood into the legal system should be taken as hemorrhage rats model method is simple,easy to operate,and is similar to the pathologic changes of the human body spontaneous cerebral hemorrhage clinical process,which is the study of the ideal animal model of cerebral hemorrhage;2.synaptic plasticity protein SYP,PSD-95 content increasing in the peripheral nerve around the hematoma after intracerebral hemorrhage,conducive to neural functional recovery;3.Thrombin can promote synaptic plasticity in the peripheral nerve around the hematoma,promote neural functional recovery after intracerebral hemorrhage.The right amount of thrombin is more advantageous to neural functional recovery after cerebral hemorrhage;4.thrombin on synaptic plasticity may be mediated through PAR1 way in the hematoma peripheral nerve;...